Objective To observe the effects of Thymosin α1 （Tα1） on acute rejection after liver transplantation and immune function of T cells. Methods Twenty recipients of liver transplantation due to primary hepatic carcinoma were divided into two groups: Tα1 group (n=10) and control group (n=10). Tα1 group received subcutaneous injection of Tα1 1.6 mg on the first day after liver transplantation and then twice a week for at least one month. Both Tα1 group and control group took same immunodepressants. Core biopsies were carried to compare the incidence rate of acute rejection between Tα1 group and control group. Peripheral T cellular immune function in these two groups was detected on 1 d before, 1 week, 2 weeks and 1 month after transplantation. Results There was not significant difference of incidence rate of acute rejection between Tα1 group and control group (Pgt;0.05). In the Tα1 group, CD4+, CD8+ lymphocyte cell counts and the CD4+/CD8+ ratio were significantly higher than those in the control group in 2 weeks and 1 month after transplantation (P＜0.05). Conclusion Use of Tα1 in recipients who also takes rountine immunosuppressants dose not increase the risk of occurring acute rejection after liver transplantation. Tα1 can significantly increase CD4+, CD8+ counts and CD4+/CD8+ ratio, which shows that Tα1 may improve recipients’ cellular immune function.
To search for the relationship between immune state and tumor metastases, CD3,CD4,CD8 and CD44 contents in 13 speciments of fine needle aspiration (FNA) from thyroid cancer patients were detected by the flowcytometry (FCM) and comparison between thyroid cancer and benign tumor was made. The result showed :in thyroid cancer group, CD＋3，CD＋4 cells and the ratio of CD＋4/CD＋8 reduced significantly (Ｐ＜0.01),and CD＋8 cell increased significantly (Ｐ＜0.01), in metastases group,this change was much significantly. CD44 expressed significantly higher in cancer group than that of the benign thyroid neoplasms, and the change was related to the tumor metastases. The results indicate that CD44 can be as a marker of tumor and indicator of metastases. The disturbance of immune system results in active expression of CD44 by tumor cells, so, lead to metastases. It is helpful to the diagnosis of thyroid cancer, assessment of metastases and management in surgery.
Objective To explore the effect of CD3AK cells on T lymphocyte subsets and its functions of patients with primary liver carcinoma (PLC). MethodsFiftyeight patients with PLC were divided into two groups, CD3AK group (n=37), control group (n=21). Eleven blood donors were taken as normal control. All patients from the treatment group were given 2×109 CD3AK cells once a day for 5 days on the 1st day after operation, having received IL2 at a dose of 100 units per 24 hour by intravenous injection starting 30 minutes before administration of CD3AK cells. T lymphocyte subsets, IL2R, NK activity, LAK activity and proliferative activity were determined. ResultsThe CD4/CD8 ratio, expression of IL2R, proliferative activity, NK activity and LAK activity of T lymphocytes from the treatment group were significantly higher than those of control respectively.Conclusion The adoptive transfer of CD3AK cells could improve the disorder of T lymphocyte subsets and its functions and provide the patients with PLC with fresh abundant effectors against tumor.
Objective To investigate the expression and clinical significance of T lymphocyte subsets, natural killer (NK) cells and CD19+ B cells in the elderly with primary immune thrombocytopenia (ITP) before and after treatment. Methods The elderly ITP patients diagnosed and treated in the Songjiang Hospital Affiliated to Shanghai Jiaotong University School of Medicine (preparatory stage) between January 2014 and June 2019 were retrospectively selected as the observation group. The healthy elderly in the same period were selected as the control group. According to the treatment, the observation group was divided into effective group and ineffective group. The expression levels of T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+), NK cells and CD19+ B cells were observed and analyzed. Results A total of 75 subjects were included, including 35 in the observation group and 40 in the control group. The total effective rate was 85.71% (30/35). Before treatment, the expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the observation group were lower than those in the control group (P<0.05). There was no significant difference in other indexes between the two groups (P>0.05). After treatment, except for CD8+, the expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the observation group were higher than those before treatment (P<0.05). The expression levels of NK cells and CD19+ B cells were lower than those before treatment (P<0.05). The expression levels of T lymphocyte subsets (CD3+, CD4+ and CD4+/CD8+) in the effective group were higher than those before treatment (P<0.05), while the expression level of CD19+ B cells was lower than that before treatment (P<0.05). There was no significant difference in other indexes before and after treatment (P>0.05). There was no significant difference in the expression levels of T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+), NK cells and CD19+ B cells in the ineffective group before and after treatment (P>0.05). Conclusions T lymphocyte subsets are abnormal in elderly ITP patients. The immune abnormality of T lymphocyte may be one of the reasons for elderly patients with ITP. With the improvement of therapeutic effect, immune cell subsets have also been improved.
Objective To study the immunological tolerance induced by blocking the second signal of T cell with extrinsic cytotoxic T lymphocyteassociated antigen 4 immuno globlin(CTLA4-Ig). Methods Fifty-four BALB/C mice, inbred strains, were employed as recipients of bone allografts, using a model of heterotopic muscle pouch. The 54 mice were divided into 3 groups and18 for each group. The first group, in which the donor was C57BL/6 with intraperitoneal injection ofL6(as a control), was named AL group. The second group,also C57BL/6 with injection CTLA4-Ig, was named AC group. The third group,homologous BALB/C with injection of PBS buffer solution, was named AB group.The serum antibody, lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor, the analysis of lymphocyte subsets, a regraft experiment and histology were determined 2, 4 and 6 weeks after transplantation. The second transplantation was to regraft C57BL/6(BC group) and C3H(BHgroup) mice respectively after first 12 mice being transplantated with C57BL/6 and injected with CTLA4-Ig as to detect donor-specificity of immunological tolerance. Results Compared with AB group, AL group created more intensive immune rejection: CD4 T cell subsets(Plt;0.05), the serum antibody(Plt;0.05) and lymphocyteproliferation of the second stimulation by splenic cell and bone supernatant ofdonor (Plt;0.01 and 0.05) were significantly increased. However, the results of AC group showed that CTLA4-Ig significantly inhibited the immune rejection: CD4T cell subsets(Pgt;0.05), the serum antibody (Pgt;0.05), and lymphocyte proliferation of the second stimulation(Pgt;0.05) were similar to those of AB group. Histological observation of AC group showed that lymphocyte infiltration disappeared,cartilage and new bone formed, and bone marrow cavities emerged. A regraft experiment showed that CD4 T cell subsets (Plt;0.05) and lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor(Plt;0.05), BC group was significantly lower than those of BH group. So theimmunological tolerance induced by CTLA4-Ig was of donor-specificity. Conclusion The immunological tolerance induced by CTLA4-Ig was prolonged for 6 weeks. This study provides a brand-new path for bone transplantation, which can be helpful to other organ transplantation.
Abstract: Objective To investigate the influence of cardiopul monary bypass(CPB) to the cellular immune function of T lymphocyte. Me th ods Among 500 patients operated from March 2006 to September 2006，30 patients with rheumatic heart disease were selected randomly as the CPB group, which would replace mitral valve; 30 patients with congenital patent ductus arte reriosus as the nonCPB group, which would ligate ductus arteriosus without CPB . The blood was sampled before operation, at the end of CPB or operation, and 24 hours after operation. After T lymphocyte was seperated, the quantum o f T lymphocyte, apoptosis of T lymphocyte, ability of T lymphocyte to kill tumou r cell were measured. Results The quantum of T lymphocyte i n CPB group at the end of CPB was decreased than that before operation （50.9% ±6.8% vs. 58.5%± 9.1%,Plt;0.05）; apoptosis of T lymphocyte at the end of CPB and 24 hou rs after operation were increased than that before operation （6.5%±2.2% vs. 0. 9%±1.1%, 5.6%±1.8% vs. 0.9%±1.1%;Plt;0.01）; ability to kill tumour cell b reakdown in CPB group at the end of CPB and 24 hours after operation was decrea sed than that before operation （30.4%±6.0% vs. 37.3%±8.6%, 29.0%±4.9% vs . 37 .3%±8.6%;Plt;0.05）. Ability to kill tumour cell breakdown in CPB group was lower than that in nonCPB group at the end of CPB （30.4%±6.0% vs. 33.6%±5. 3%, Plt;0.05）. Conclusion CPB can depress the cellular im mune function,which causes temporary immune depression to the body.
ObjectiveTo investigate the method for generating anchor chemric T lymphocytes that can target tumor associated glycoprotein-72 (TAG72) antigen and analyze their repressive effects on proliferation of TAG72 positive hepatocarcinoma cells. MethodsFirstly, peripheral blood mononuclear cells (PBMCs) from healthy volunteers were isolated. And then, CD8+ T cells were isolated from PBMCs via magnetic activated cell sorting (MACS). These lymphocytes were transfected with recombinant vector, anti-TAG72-scFv-CD28-pcDNA3, through Lipofectamine2000 to gernerate anchor chimeric TAG72-specific CD8+ T cells. SMMC7721 (TAG72 positive) hepatocarcinoma cells were co-cultured with chimeric T lymphocytes and their cell cycles were analyzed by flow cytometry (FCM). ResultsAnchor chmeric T lymphcytes targetting TAG72 recognized TAG72 positive SMM7721 cells and repressive effects on their proliferation were observed by flow cytometry. ConclusionAnchor chmeric T lymphcytes targetting TAG72 on tumor surface can specifically recognize TAG72 positive hepatocarcinoma cells and may exert repressive effect on their proliferation.
Objective To investigate the effects of down-regulating of Rfng gene ( 1, 3-Nacetylglucosaminyltransferases) in lung CD4 + T cells of asthmatic rat model by small interfering RNA ( siRNA) and explore the role of Rfng in pathogenesis of asthma. Methods An asthmatic rat model was established by OVA sensitization and challenge. Total T cells were isolated from lung tissue of asthmatic rats, and CD4 + T lymphocytes were purified using magnetic beads. CD4 + T lymphocytes were transfected by siRNA targeting Rfng gene. The mRNA and protein expressions of Rfng were detected by quantitative PCR and Western blot. Quantitative PCR was performed to determine the mRNA levels of Th1 /Th2 cytokines and related genes including IL-12, IFN-γ, IL-4, IL-5, T-bet, and GATA3. ELISA was performed to determine the concentrations of IL-12, IFN-γ, IL-4, and IL-5 in supernatant. Results The mRNA and protein expression of Rfng in RNAi group decreased significantly. IL-12, IFN-γ, T-bet increased and while IL-4, IL-5, and GATA3 decreased significantly. The concentrations of IL-12 and IFN-γ in the supernatant increased significantly, while IL-4 and IL-5 decreased significantly. Conclusions Down regulation of Rfng affects T cell differentiation. It is presumed that Fringe contribute to the pathogenesis of asthma.
【Abstract】Objective This study was conducted to build experimental model of orthotopic liver transplantation in rat (ROLT) with the character of acute rejection; and to study the effect of cytotoxic T lymphocyte antigen 4 immunoglobulin G (CTLA4Ig) on prevention of rejection and the induction of immune tolerance of ROLT. Methods Build model of Wistar→SD ROLT(performed by the two cuff method) with character of acute rejection.Recipients were injected with CTLA4Ig 75 μg per ROLT into abdominal cavity after 2 days of operation. Contrast was made with no treatment group, the clinical characters, the liver function, the transplantated liver pathologic character and the concentrations of TNFα in serum were observed and measured on postoperative day 7. In treatment group, all above observation were done on postoperative month 4. Above all, determination of the effect of CTLA4Ig on preventing acute rejection and inducing tolerance in ROLT was observed.Results ①Recipients (no treatment group) died one by one within 6th～14th days; pathologic character of rejection in transplantation liver could be found; ② In treatment group, on postoperative day 7 and month 4, no clinical rejection character and no pathologic character of rejection in transplantation liver were found and serum concentration of cytokins related to TNFα found lower than that of contrast group(P＜0.05), and serum concentration of ALT、AST、TBIL、DBIL found lower too(P＜0.05); But serum concentration of TP and Alb was found higher than that of contrast group(P＜0.05). Conclusion ① CTLA4Ig treatment alone inhibits the rejection responce in ROLT. ② CTLA4Ig treatment can Prevent rejection and induce immune tolerance in ROLT model with characters of acute rejection; the serum concentration of cytokins related to TNFα can probably be used as evaluation standard of rejection in ROLT rejection.
Objective To study effect of carcinoembryonic antigen (CEA) positive targeted lymphocytes on gastric cancer cells in vitro and in vivo. Methods The peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers. The recombinant vector anti-CEA-scFv-CD3ζ-pcDNA3.0 was transfected into the PBMCs by lipofectamine 2000, by this means, the CEA special lymphocytes were obtained. Meanwhile, the PBMCs transfected with empty plasmid pcDNA3.0 were used as control (empty vector lymphocytes). The different lymphocytes and gastric cancer cells (CEA positive KATOⅢ gastric cancer cells and CEA negative BGC-823 gastric cancer cells) were co-cultured, then the ability to identify the gastric cancer cells and it’s effect on apoptosis of gastric cancer cells were observed at 24 h or 36 h later respectively. The CEA special lymphocytes and empty vector lymphocytes were injected by the tail vein of nude mice bearing gastric cancer cells, then it’s effect on the tumor was observed. Results ① The CEA special lymphocytes could strongly identify the KATOⅢ gastric cancer cells (identification rate was 72.3%), which could weakly identify the BGC-823 gastric cancer cells (identification rate was 7.8%). ② The apoptosis rate of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly higher than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (P=0.032), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (P=0.118). ③ The tumor volume of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly smaller than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (F=5.010, P<0.01) or the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells (F=4.982, P<0.01), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (F=1.210, P>0.05). Conclusion CEA special lymphocytes can promote cell apoptosis and inhabit tumor reproduction of CEA positive gastric cancer cells in vitro and in vivo.